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4种中草药单体对嗜热四膜虫生长和凋亡的影响

潘厚军 康佳磊 张德锋 常藕琴 任燕 王亚军 蒋俊贤 王琳 石存斌

潘厚军, 康佳磊, 张德锋, 常藕琴, 任燕, 王亚军, 蒋俊贤, 王琳, 石存斌. 4种中草药单体对嗜热四膜虫生长和凋亡的影响[J]. 南方水产科学. doi: 10.12131/20210163
引用本文: 潘厚军, 康佳磊, 张德锋, 常藕琴, 任燕, 王亚军, 蒋俊贤, 王琳, 石存斌. 4种中草药单体对嗜热四膜虫生长和凋亡的影响[J]. 南方水产科学. doi: 10.12131/20210163
PAN Houjun, KANG Jialei, ZHANG Defeng, CHANG Ouqin, REN Yan, WANG Yajun, JIANG Junxian, WANG Lin, SHI Cunbin. Effects of four kinds of Chinese herbal monomers on growth and apoptosis of Tetrahymena thermophila[J]. South China Fisheries Science. doi: 10.12131/20210163
Citation: PAN Houjun, KANG Jialei, ZHANG Defeng, CHANG Ouqin, REN Yan, WANG Yajun, JIANG Junxian, WANG Lin, SHI Cunbin. Effects of four kinds of Chinese herbal monomers on growth and apoptosis of Tetrahymena thermophila[J]. South China Fisheries Science. doi: 10.12131/20210163

4种中草药单体对嗜热四膜虫生长和凋亡的影响

doi: 10.12131/20210163
基金项目: 广州市科技计划项目 (201803020045);财政部和农业农村部国家现代农业产业技术体系 (CARS-45);广东省自然科学基金项目 (2017A030313172)
详细信息
    作者简介:

    潘厚军 (1968—),女,硕士,研究员,从事鱼类病害与防治技术研究。E-mail: phj001@126.com

    通讯作者:

    石存斌 (1964—),男,硕士,研究员,从事鱼类病害与防治技术研究。E-mail: shicunbin2006@163.com

  • 中图分类号: S 948

Effects of four kinds of Chinese herbal monomers on growth and apoptosis of Tetrahymena thermophila

  • 摘要: 利用Hoechst 33258荧光染色、活细胞计数、CCK-8细胞活力测定、抗氧化酶活力测定等技术,研究了青蒿素、大黄素、芦荟大黄素、小檗碱 (又名黄连素) 等4种中草药单体对嗜热四膜虫 (Tetrahymena thermophila) 生长和凋亡的影响,为应用嗜热四膜虫筛选抗鱼寄生虫中草药提供参考。结果表明,采用Hoechst荧光染色,4种单体在高浓度 (20~200 mg·L−1) 作用下均可使嗜热四膜虫凋亡,显著着色发亮。4种单体对嗜热四膜虫的半抑制浓度 (IC50) 和95%置信区间 (95% confidence interval, 95% CI),用细胞计数法测定依次为21.23 (16.71~28.23) mg·L−1、3.77 (1.91~5.54) mg·L−1、1.70 (0.18~3.82) mg·L−1和44.56 (29.82~98.66) mg·L−1,CCK-8细胞活力测定法依次为20.03 (13.02~28.45) mg·L−1、6.07 (4.39~7.65) mg·L−1、3.67 (2.61~4.72) mg·L−1和107.39 (66.78~267.17) mg·L−1,CCK-8法与细胞计数法所呈现的趋势较为一致。在单体质量浓度为1.25~50.0 mg·L−1,随着药物浓度的增加,过氧化氢酶 (CAT) 活力有升高趋势,而丙二醛 (MDA) 呈下降趋势,超氧化物歧化酶 (SOD) 活力先升高后降低。上述结果表明,4种中草药单体对嗜热四膜虫的生长和凋亡有剂量效应关系,凋亡的产生可能与细胞氧化应激状态改变有关。
  • 图  1  4种中草药单体对嗜热四膜虫Cu428.2凋亡的影响 (Hoechst 33258试剂染色)

    A. 青蒿素;B. 大黄素;C. 芦荟大黄素;D. 小檗碱;1、2、3、4示药物质量浓度分别为0、2.0、20、200 mg·L−1

    Figure  1.  Effect of four kinds of Chinese herbal monomers on apoptotic morphology of T. thermophila Cu428.2 (stained by Hoechst 33258)

    A. Artemisinin; B. Emodin; C. Aloe-emodin; D. Berberine; 1, 2, 3 and 4 show the monomer concentration as 0, 2.0, 20, 200 mg·L−1; respectively.

    图  2  不同浓度的4种单体作用下嗜热四膜虫Cu428.2株的活细胞密度和CCK-8细胞活力

    a. 活细胞密度;b. CCK-8细胞活力。

    Figure  2.  Cell Density and CCK-8 Cell viability of T. thermophila Cu428.2 at different concentrations of four kinds of Chinese herbal monomers

    a. Living cell density; b. CCK-8 cell viability.

    图  3  嗜热四膜虫Cu428.2在4种中草药单体分别处理后超氧化物歧化酶、过氧化氢酶、丙二醛活力

    *. 与对照组差异显著 (P<0.05);**. 与对照组差异极显著 (P<0.01)。

    Figure  3.  SOD, CAT and MDA activity of T. thermophila Cu428.2 after treatment of four kinds of Chinese herbal monomers

    *. The difference is significant with the control group (P<0.05); **. The difference is extraordinary significant with the control group (P<0.01).

    表  1  实验的4种中草药单体

    Table  1.   Four kinds of Chinese herbal monomers used in experiment

    中草药单体
    Chinese herbal monomer
    所属类别
    Classification
    来源厂家
    Resource
    质量分数
    Mass fraction/%
    青蒿素 Artemisinin 倍半萜内酯类 大连美仑 97
    大黄素 Emodin 蒽醌类 大连美仑 98
    芦荟大黄素 Aloe-emodin 蒽醌类 大连美仑 96
    小檗碱 Berberine 季胺生物碱类 大连美仑 98
    下载: 导出CSV

    表  2  活细胞计数法、CCK-8法测得的4种中草药单体对嗜热四膜虫的半抑制浓度

    Table  2.   50% inhibitory concentration (IC50) of four kinds of Chinese herbal monomers to T.thermophila Cu428.2 determined by living cell counting method and CCK-8 test

    中草药单体
    Chinese herbal monomer
    活细胞计数法
    Living cell counting method/(mg·L−1)
    CCK-8法
    CCK-8 test/(mg·L−1)
    半抑制浓度
    IC50
    95%置信区间
    95% CI
    半抑制浓度
    IC50
    95%置信区间
    95% CI
    青蒿素 Artemisinin 21.23 16.71~28.23 20.03 13.02~28.45
    大黄素 Emodin 3.77 1.91~5.54 6.07 4.39~7.65
    芦荟大黄素 Aloe-emodin 1.70 0.18~3.82 3.67 2.61~4.72
    小檗碱 Berberine 44.56 29.82~98.66 107.39 66.78~267.17
    下载: 导出CSV
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  • 收稿日期:  2021-06-02
  • 修回日期:  2021-09-08
  • 录用日期:  2021-09-28
  • 网络出版日期:  2021-10-19

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